This SBIR project is focused to develop, validate and commercialize an advanced high-throughput anti- tumor drug screening platform (HTS) which employs a new 3D HuBioGEM assay technology. Preclinical drug screening is commonly performed using biochemical and cell-based assay systems. In recent years, more relevant 3D spheroid models have been developed using microcarrier, pHEMA- coating, gel-layer and hanging-drop techniques. Although useful, these require difficult culture protocols, provide undesirable endpoints and/or are not efficient for automated applications. We have created a novel HuBioGEM culture system by formulating patented HuBiogel (human biomatrix) with unique paramagnetic particles (GEM). Employing 3D magnetic-biogel and 384-array culture strategies, we propose to develop a robust human microtumor assay platform for drug efficacy profiling. Phase I technical goals are: i) Establish 3D tumor production, culture and analysis protocols using HuBioGEM, NCI cell lines and custom array tools; and ii) Demonstrate utility of HTS assay system for evaluating anti-tumor activity of many known drugs. Our integrated approach will allow automated handling, imaging and functional analysis. New 3D tumor system will be compared with current spheroid model to confirm its practical advantages. We are confident this 3D tumor biology platform will permit robust and accurate prediction of preclinical drug candidates and thus will accelerate drug discovery and development programs. Commercial Importance & Significance: No satisfactory HTS assay system currently exists in market which utilizes fully-human 3D cell culture technology. New 384-well tumor bioassay will be useful for automated imaging, functional, proteomic and genomic analysis platforms. Sales of HuBioGEM culture kits and Ready-use preclinical assay screens would have significant world-wide market.